The Michael Hooker Microscopy Core Facility

 

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The Michael Hooker Microscopy Facility (MHMF) is a research light microscopy facility providing advanced digital light microscopy, image processing and analysis resources for users from the UNC Chapel Hill campus. We offer instrumentation and instruction to enable users to acquire, process and analyze images from a wide variety of sample types.


Equipment Available

New! Olympus FV1000 MPE SIM Laser Scanning Confocal Microscope


MHMF-1

  • Laser Spot Scanning Confocal Microscope
  • Inverted microscope stand with widefield DIC
  • Visible excitation lasers switched/modulated using an AOTF
  • Multiphoton excitation laser modulated with an AOM
  • Simultaneous transmitted light or DIC imaging while scanning confocally (single or two photon)
  • Three standard Photo Multipliers Tube (PMT) light detectors with 2 spectral discriminations
  • Motorized x-y stage- tilting, tilting with time lapse
  • Environmental chamber for controlled temperature, gas, and humidity
  • FRAP/FLIP/FLIM/FRET/PhotoActivation/etc. - SIM scanner
  • True 12 bit acquisition- 4096 pixel grey level

Detection:

  • Filtering through glass interference filters
  • Spectral separation through diffraction gratings
  • Two tunable detection bands (1-100 nm band width) with adjustable spectral windows before the three Photo Multiplier Tubes (PMT)
  • Standard XY scanning with zoom and rotation
  • Transmitted light and DIC (non confocal)
  • FRAP, FRET, photoactivation, Emission Spectra, ratio imaging
  • Two Channel non descanned detectors (NDD)

Objectives:

Mag.NATypeWDCorrectionsCover SlipImmersion
10x 0.4 UPlansaPO 3.1 mm #1.5 air
20x 0.75 UPlanSApo 550 um #1.5 air
30x 1.05 UPlanSApo 800 um corr 0.13-0.19 silicon oil
60x 1.42 PlanApo 150 um #1.5

oil


More information and how to get started using the Olympus FV1000


Advanced and Standard Microscopy

  • Standard transmitted color photomicrographs
  • Fluorescence photomicrographs
  • Phase contrast and DIC/Nomarski
  • Ratio imaging. e.g. Ca2+ Fura-2
  • Time-lapse (fluorescence and transmitted light)
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Airway Epithelium, (Dr. Silvia Kreda). H & E airway epithelium.

 

Additional Confocal Microscopy

  • Multichannel fluorescence + DIC overlayed
  • Fast X-Z scanning 3D and 4D image acquisition
  • Spectral Imaging - dye emission unmixing
  • Co-localization & analysis
  • Time lapse
  • Fluorescence Recovery After Photobleaching (FRAP)
  • Fluorescence Resonance Energy Transfer (FRET)
  • Deconvolution
MHM14.jpg Airway epithelia. Green: NaK-ATPase. Red: Propidium Iodide. (Dr. Alessandra Livraghi).

 

Live Cell Imaging

  • 3-color Spinning-disc Confocal (FITC, Texas Red & CY5 like dyes)
  • Simultaneous fluorescence and transmitted light (DIC)
  • Heated stages, chambers, objective heater
  • Temperature and environmental control

Color Brightfield Imaging (high resolution)

  • Nikon DXM 1200 pixel shift color camera (12 MegaPixels)
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    Scattergram showing no co-localization between the red & green antibody labels, Dr. Karen Loechner. Mouse Embryo 3D render, Dr. Jaime Rivera.

Stereo Fluorescence Micro/Macroscope

  • Fluorescence/standard illumination
  • Motorized focus & filter changers
  • Automated acquisition – z-series/time lapse/multi channel

Laser Micro-Dissection System

  • Isolation of subcellular and tissue regions
  • For DNA/RNA amplification or protein for mass spectroscopy
  • Cauterization of vessel in developing organism

Image Processing and Analysis

  • Co-localization analysis
  • 3-D Image rendering and quantification
  • Particle tracking – 2D & 3D
  • Fluorescence quantification
  • Densitometry
  • Area/volume measurements
  • C-Imaging, Metamorph (offline)
  • Volocity – Decovolution, 3D rendering, 3D quantification, colocalization, tracking
  • Volocity remote access license server

http://microscopy.unc.edu/volocity

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Airway Epithelium, Dr. Raymond Pickles

"Brainbow" Dr. Alain Burette, Cell & Devel. Biology

 

Other Resources

 

MHMF Personnel and Contact Information

MHM17.jpg Dr. Michael Chua
Cell & Molecular Physiology
6007 Thurston Bowles
919-843-3268
microscopy@unc.edu
MHM18.jpg Dr. Neal Kramarcy
6129 Thurston Bowles
919-966-7051
kramarcy@med.unc.edu

 

 

 

 

 

 

 

 


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